Enzymatic Synthesis of N-Acetyllactosamine (LacNAc) Type 1 Oligomers and Characterization as Multivalent Galectin Ligands.
Enzymatic Synthesis of N-Acetyllactosamine (LacNAc) Type 1 Oligomers and Characterization as Multivalent Galectin Ligands.
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DOI：
10.3390/molecules22081320
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发表时间：
2017-08-10
期刊：
影响因子：
--
通讯作者：
Elling L
中科院分区：
文献类型：
--
作者：
Fischöder T;Laaf D;Dey C;Elling L
二糖单位N-乙酰乳糖胺(LacNAc)的重复序列在糖蛋白和糖脂上以1型(Gal1β1，3GlcNAc)和2型(Gal1β1，4GlcNAc)糖基化基序的形式出现。LacNAc基序作为凝集素的结合配体，参与了许多生物识别事件。据我们所知，我们首次利用来自大肠杆菌的重组β1，3-半乳糖基转移酶和来自幽门螺杆菌的β1，3-N-乙酰氨基葡萄糖转移酶合成了LacNAc 1型寡聚体。在还原或非还原末端分别呈现LacNAc-1重复序列或LacNAc-1的四糖多糖通过方酸连接物化学连接到牛血清白蛋白上作为蛋白质支架。在酶联凝集素试验(ELLA)中，进一步研究了呈现新糖蛋白的多价LacNAc 1型新糖蛋白与肿瘤相关的人半乳糖凝集素3(Gal-3)及其截短的对应物Gal-3∆的特异性结合。我们观察到Gal-3∆对呈现LacNAc类型1重复单位的多价新糖蛋白的亲和力显著增加。这是Gal-3、∆和Gal-3在糖链选择性上存在差异的第一个证据，并可能进一步用于在肿瘤进展和治疗过程中追踪Gal-3∆。
Repeats of the disaccharide unit N-acetyllactosamine (LacNAc) occur as type 1 (Galβ1, 3GlcNAc) and type 2 (Galβ1, 4GlcNAc) glycosylation motifs on glycoproteins and glycolipids. The LacNAc motif acts as binding ligand for lectins and is involved in many biological recognition events. To the best of our knowledge, we present, for the first time, the synthesis of LacNAc type 1 oligomers using recombinant β1,3-galactosyltransferase from Escherichia coli and β1,3-N-acetylglucosaminyltranferase from Helicobacter pylori. Tetrasaccharide glycans presenting LacNAc type 1 repeats or LacNAc type 1 at the reducing or non-reducing end, respectively, were conjugated to bovine serum albumin as a protein scaffold by squarate linker chemistry. The resulting multivalent LacNAc type 1 presenting neo-glycoproteins were further studied for specific binding of the tumor-associated human galectin 3 (Gal-3) and its truncated counterpart Gal-3∆ in an enzyme-linked lectin assay (ELLA). We observed a significantly increased affinity of Gal-3∆ towards the multivalent neo-glycoprotein presenting LacNAc type 1 repeating units. This is the first evidence for differences in glycan selectivity of Gal-3∆ and Gal-3 and may be further utilized for tracing Gal-3∆ during tumor progression and therapy.
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DOI：
10.1074/jbc.m116.768887
发表时间：
2017-04-14
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Choo M;Tan HL;Ding V;Castangia R;Belgacem O;Liau B;Hartley-Tassell L;Haslam SM;Dell A;Choo A
通讯作者：
Choo A
影响因子：
27.4
作者：
Guévremont, M;Martel-Pelletier, J;Reboul, P
通讯作者：
Reboul, P
DOI：
10.1016/0006-291x(83)91160-9
发表时间：
1983-01-01
影响因子：
3.1
作者：
FALK, KE;KARLSSON, KA;KOPROWSKI, H
通讯作者：
KOPROWSKI, H
影响因子：
4.9
作者：
Chen, Yi;Thon, Vireak;Chen, Xi
通讯作者：
Chen, Xi
影响因子：
4.3
作者：
Boecker, Sophia;Elling, Lothar
通讯作者：
Elling, Lothar